If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO340
Recent progress of this line F2 individuals have been indentified
Gene name si:dkey-256e7.8
Ensembl ID ENSDARG00000092022
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGAAAGAGTTCAGACAGAGCtgg
Cas9 target site 2 sequence
Position in the gene Exon 2
Restriction enzyme (RE) Hpy 188I(TCN|GA)
PCR primer (forward) GAATCACCCAAAGCAAACA
PCR primer (reverse) TTACACGGAGCTTACCTTCA
Length of expected PCR product (bp) 431
Length of expected RE digestion product (bp) 76+138+217
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 340
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 30
Survival rate of F0 embryo @ 1dpf (%) 74
Method to estimate mutation efficiency Restriction enzyme digestionRE digestion (Hpy 188I)
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO340.pdf
Mutation efficiency in F0 embryos (%) 20
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 28%(2/7)
Wild-type (WT) sequence GGAAAGAGTTCAGACAGAGC…CTCTTAA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GGAAAGAGTTCAGACAG--...--CTCTTAA(-179) pcr直接鉴定
F0-2 GGAAAGAGTTCAGAC----------GAAGT(-10) 需要测序鉴定
Allele name and predicted product 340a
340b
GGAAAGAGTTCAGACAG--...--CTCTTAA; Truncated protein(249aa)
GGAAAGAGTTCAGAC----------GAAGT; Truncated protein(257aa)
突变创建时间 2014-04-10
填表时间 2013/12/18
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information: